Journal: Journal of Translational Medicine

Article Title: IκBα kinase inhibitor BAY 11-7082 promotes anti-tumor effect in RAS-driven cancers

doi: 10.1186/s12967-024-05384-4

Figure Lengend Snippet: BAY 11-7082 treatment alters the expression of multiple genes in NRAS, KRAS, and HRAS mutant cancer cells. (A) Heatmaps showing the top 50 upregulated or downregulated genes in SKMEL-103, AsPC1, and RH-36 cells upon treatment with BAY 11-7082 (5 µM) for 48 h as compared with DMSO-treated cells. (B) Volcano plot showing top 15 genes upregulated or downregulated after 48-h treatment with BAY 11-7082 (5 µM) in RAS mutant cells

Article Snippet: RAS mutant cancer cell lines (NRAS: M245, SKMEL-103, SKMEL-2; KRAS: PANC1, AsPC1, SU.86.86) were purchased from American Type Culture Collection (ATCC) as listed in Supplementary Tables and maintained in a humidified atmosphere of 5% CO2 at 37 °C in Dulbecco’s modified Eagle medium (Life Technologies, Carlsbad, CA, USA) or Roswell Park Memorial Institute-1640 Medium (Life Technologies), each supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (both from Life Technologies).

Techniques: Expressing, Mutagenesis

Journal: Journal of Translational Medicine

Article Title: IκBα kinase inhibitor BAY 11-7082 promotes anti-tumor effect in RAS-driven cancers

doi: 10.1186/s12967-024-05384-4

Figure Lengend Snippet: Model showing the new IκBα kinase Inhibitor BAY 11-7082 for Treating RAS-driven cancers. A model depicting various cancer types driven by oncogenic NRAS, KRAS, and HRAS mutations overexpress IκBα kinase. IκBα kinase can be effectively targeted by small molecule inhibitor BAY 11-7082, leading to downregulation of several prooncogenic signaling pathways, including PI3K-AKT signaling cascade and upregulation of apoptosis that contributes to tumor growth inhibition observed in RAS-driven cancers

Article Snippet: RAS mutant cancer cell lines (NRAS: M245, SKMEL-103, SKMEL-2; KRAS: PANC1, AsPC1, SU.86.86) were purchased from American Type Culture Collection (ATCC) as listed in Supplementary Tables and maintained in a humidified atmosphere of 5% CO2 at 37 °C in Dulbecco’s modified Eagle medium (Life Technologies, Carlsbad, CA, USA) or Roswell Park Memorial Institute-1640 Medium (Life Technologies), each supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin (both from Life Technologies).

Techniques: Protein-Protein interactions, Inhibition

Journal: Cell systems

Article Title: Dissecting RAF inhibitor resistance by structure-based modeling reveals ways to overcome oncogenic RAS signaling

doi: 10.1016/j.cels.2018.06.002

Figure Lengend Snippet: Key resources table

Article Snippet: Reagent or resource Source Identifier Antibodies Anti-MAP Kinase (ERK-1, ERK-2) antibody produced in rabbit Sigma-Aldrich Cat. No. M5670 Monoclonal Anti-MAP Kinase, Activated (Diphosphorylated ERK-1&2) antibody produced in mouse Sigma-Aldrich Cat. No. M8159 Anti-rabbit IgG, HRP-linked Antibody Cell Signalling Technologies Cat. No. 7074 Anti-mouse IgG, HRP-linked Antibody Cell Signalling Technologies Cat. No. 7076 Chemicals, Peptides, and Recombinant Proteins Vemurafenib (PLX4032) Selleckchem Cat. No. S1267 Dabrafenib (GSK2118436) Selleckchem Cat. No.S2807 TAK-632 Selleckchem Cat. No.S7291 Trametinib (GSK1120212) Selleckchem Cat. No.S2673 Sorafenib tosylate Axon Medchem Axon 1397 SB-590885 R&D Systems 2650/10 B0R (2,6-difluoro-N-(3-methoxy-2H-pyrazolo[3,4-b]pyridin-5-yl)-3-[(propylsulfonyl) amino]benzamide) Genentech, ( Wenglowsky et al., 2011 ) B0R Critical Commercial Assays pMEK assay ProtATonce P-MP2K1-A01 pERK assay ProtATonce P-MK03-A01 GAPDH assay ProtATonce P-GAPDH-A01 Assay buffer ProtATonce PPK40-A01 Lysis buffer ProtATonce PLY40-A01 Phospho(Ser217/221)/Total MEK1/2 Assay Whole Cell Lysate Kit Meso Scale Discovery K15129D Phospho(Thr202/Tyr204; Thr185/Tyr187)/Total ERK1/2 Assay Whole Cell Lysate Kit Meso Scale Discovery K15107D PierceTM BCA Protein Assay Kit ThermoFisher Scientific/Pierce Cat. No. 23225 CellTiter 96® Aqueous One Solution Cell Proliferation Assay (MTS) Promega Cat. No. G3581 Deposited Data Dose-response MESOSCALE data of MEK and ERK signaling in cells treated with RAF inhibitors This study Data S4.xlsx Dose-response Luminex data of MEK and ERK signaling in cells treated with RAF inhibitors This study Data S4.xlsx Experimental Models: Cell Lines SKMEL2 ATCC HTB-68 A375 ATCC CRL-1619 MEL-JUSO DSMZ ACC 74 M249 parental and resistant Antoni Ribas ( Nazarian et al., 2010 ) Software and Algorithms Calculation number of colonies Clono-Counter ( Niyazi et al., 2007 ) Simulation of rule-based models BioNetGen http://www.csb.pitt.edu/Faculty/Faeder/?page_id=409 Spreadsheet for calculation of apparent dissociation constants.

Techniques: Produced, Recombinant, Lysis, Bicinchoninic Acid Protein Assay, Proliferation Assay, Luminex, Software